Kim J, Hirsch J P
Department of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029, USA.
Genetics. 1998 Jun;149(2):795-805. doi: 10.1093/genetics/149.2.795.
SSF1 and SSF2 are redundant essential yeast genes that, when overexpressed, increase the mating efficiency of cells containing a defective Ste4p Gbeta subunit. To identify the precise function of these genes in mating, different responses to pheromone were assayed in cells that either lacked or overexpressed SSF gene products. Cells containing null alleles of both SSF1 and SSF2 displayed the normal transcriptional induction response to pheromone but were unable to form mating projections. Overexpression of SSF1 conferred the ability to form mating projections on cells containing a temperature-sensitive STE4 allele, but had only a small effect on transcriptional induction. SSF1 overexpression preferentially increased the mating efficiency of a strain containing a null allele of SPA2, a gene that functions specifically in cell morphology. To investigate whether Ssf1p plays a direct physical role in mating projection formation, its subcellular location was determined. An Ssf1p-GFP fusion was found to localize to the nucleolus, implying that the role of SSF gene products in projection formation is indirect. The region of Ssf1p-GFP localization in cells undergoing projection formation was larger and more diffuse, and was often present in a specific orientation with respect to the projection. Although the function of Ssf1p appears to originate in the nucleus, it is likely that it ultimately acts on one or more of the proteins that is directly involved in the morphological response to pheromone. Because many of the proteins required for projection formation during mating are also required for bud formation during vegetative growth, regulation of the activity or amount of one or more of these proteins by Ssf1p could explain its role in both mating and dividing cells.
SSF1和SSF2是酵母中功能冗余的必需基因,当它们过度表达时,会提高含有缺陷型Ste4p Gβ亚基的细胞的交配效率。为了确定这些基因在交配过程中的精确功能,我们在缺乏或过度表达SSF基因产物的细胞中检测了对信息素的不同反应。同时缺失SSF1和SSF2无效等位基因的细胞对信息素表现出正常的转录诱导反应,但无法形成交配突起。SSF1的过度表达赋予了含有温度敏感型STE4等位基因的细胞形成交配突起的能力,但对转录诱导只有很小的影响。SSF1的过度表达优先提高了含有SPA2无效等位基因的菌株的交配效率,SPA2是一个专门在细胞形态中发挥作用的基因。为了研究Ssf1p在交配突起形成中是否发挥直接的物理作用,我们确定了它的亚细胞定位。发现Ssf1p-GFP融合蛋白定位于核仁,这意味着SSF基因产物在突起形成中的作用是间接的。在形成突起的细胞中,Ssf1p-GFP的定位区域更大且更弥散,并且通常相对于突起以特定方向存在。尽管Ssf1p的功能似乎起源于细胞核,但它很可能最终作用于一种或多种直接参与对信息素形态反应的蛋白质。因为交配过程中形成突起所需的许多蛋白质在营养生长过程中形成芽时也需要,所以Ssf1p对这些蛋白质中一种或多种的活性或数量的调节可以解释它在交配和分裂细胞中的作用。